Page 81 - Application Handbook - Liquid Chromatography

Application No.L463
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n Repeatability mV
Table 2 and 3 show the relative standard deviations (n=6) 1000
of retention time and peak area obtained in repeat analysis 1 n Peaks
of a mixed histamine and tyramine standard solution (each 1. Histamine
1 mg/L). Limits for histamine differ depending on the 750 2. Tyramine
country, but in Codex, for example, the limit associated
with decomposition in fish and fishery products is 500
100 mg/kg. Good repeatability has been obtained at a
concentration of about 1/100 of the criterion. 2
250
Table 2 Repeatability of Peak Table 3 Repeatability of Peak
Area and Retention Area and Retention
Time of Histamine Time of Tyramine 0
0 5 10 15 min
R.t (min) Peak Area R.t (min) Peak Area
1st 9.962 433,724 1st 11.844 153,458 Fig. 5 Chromatogram of Red Wine
2nd 9.983 431,874 2nd 11.871 155,582
3rd 9.967 441,528 3rd 11.858 155,848 mV
4th 9.962 429,887 4th 11.855 154,509 1000
5th 9.972 439,560 5th 11.882 151,206 ■Peaks
6th 9.993 434,818 6th 11.911 153,960 1 1. Histamine
Ave 9.973 435,232 Ave 11.87 154,094 750 2. Tyramine
%RSD 0.12 1.03 %RSD 0.20 1.09
500
n Analysis of Food
Figures 3 to 8 show examples of analysis of commercial 2
fish sauce, wine, and soy sauce. Pretreatment consisted 250
of preparing a 10-fold dilution using pH 2.2 sodium
citrate buffer solution, and filtering through a 0.2 µm 0
pore diameter membrane filter. As for the red wine and 0 5 10 15 min
white wine, both were spiked with histamine and
tyramine at 50 mg/L each. Fig. 6 Chromatogram of White Wine

mV mV
2000
n Peaks
500 1. Histamine n Peaks
2. Tyramine 1. Histamine
1500 2. Tyramine
1
2
1000
250
1
500
2
0
0 5 10 15 min 0 0 5 10 15 min

Fig. 3 Chromatogram of Fish Sauce A Fig. 7 Chromatogram of Soy Sauce A

mV mV
500
n Peaks n Peaks
500 1. Histamine 1. Histamine
2. Tyramine 2. Tyramine

250
250 1
2
2 1
0 0
0 5 10 15 min 0 5 10 15 min

Fig. 4 Chromatogram of Fish Sauce B Fig. 8 Chromatogram of Soy Sauce B
First Edition: Sep. 2014

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